Size exclusion chromatography of polypeptides on Bio-Gel P2 in the dynamic axial compression mode

The effect of dynamic axial compression (DAC) upon the structure of a bed packed with semirigid Bio-Gel P2 and its chromatographic parameters was studied in the separation of peptides of various mass. In the studied range of DAC pressure, 0–5 bar, a decrease in the retention times of the studied substances and the standard deviation of their peaks is observed. In spite of a reduced packed bed height and an increase in the linear velocity of the eluent, the height equivalent to the theoretical plate (HETP) does not rise. The resolution of the separated substances increases throughout the studied range of DAC pressure with the Bio-Gel P2 exclusion limit extended. According to the results obtained, DAC provides more efficient optimisation of the bed parameters in columns packed with Bio-Gel P2 than in columns packed with Sephadex G-25 due to the greater rigidity of the former packing.